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Does not react withZebrafish
Synthetic peptide derived from within residues 1 - 100 of Human beta Actin.
(Peptide available as ab13772.)
Our Abpromise guarantee covers the use of ab8227 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||1/1000 - 1/5000. Can be blocked with Human beta Actin peptide (ab13772).|
|ICC||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.|
|IHC-FrFl||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 1 µg/ml.|
IHC image of beta Actin staining in normal human colon, formalin-fixed and paraffin-embedded tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8227, 3µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab97200, 1/200 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab8227 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab8227 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Rabbit polyclonal to beta actin (ab8227) at 1/5000. Lane 1: HeLa whole cell extract (20ug); lane 2: yeast cell extract (20ug); lane 3: mouse brain tissue lysate (20ug).
Developed using Goat anti-rabbit IgG (HRP) (ab6721) at 1/2000.
ICC/IF image of ab8227 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab8227, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue).
Western blot using ab8227 at 1/1000.
Lane 1: HeLa cells (Human)
Lane 2: 3T3 cells (Mouse)
Lane 3: Fish Liver
Lane 4: Rabbit Liver
Lane 5: MDCK cells (Dog)
Lane 6: EBTr cells (Cow)
Lane 7: SL-29 cells (Chicken)
Lane 8: CHO cells (Chinese Hamster)
Lane 9: Xenopus embryo
Secondary ab: Rabbit polyclonal to Goat IgG H&L (HRP) ab6741 (1/5000)
Exposure time: 30 sec
Lysates at 20
Expected molecular weight: 41.7kDa.
ab8227 staining beta Actin in human LGE neural progenitors by Flow Cytometry. Cells were paraformaldehyde and permeabilized. The sample was incubated with the primary antibody (1/300) for 1 hour at 20°C. A PerCP-conjugated goat anti-rabbit IgG (1/800) was used as the secondary antibody.
Gating Strategy: Isotype population (shown in white).
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